Ana Carolina Acosta

Crystalline lens MTF measurement during simulated accommodation

Purpose: To design and test an optical system to measure the optical quality of post mortem lenses during simulated accommodation. Methods: An optical bench top system was designed to measure the point spread function and calculate the modulation transfer function (MTF) of monkey and human ex-vivo crystalline lenses. The system consists of a super luminescent diode emitting at 850nm, collimated into a 3mm beam which is focused by the ex-vivo lens under test. The intensity distribution at the focus (point spread function) is re-imaged and magnified onto a beam profiler CCD camera. The optical quality in terms of spatial frequency response (modulation transfer function) is calculated by Fourier transform of the point spread function. The system was used on ex-vivo lenses with attached zonules, ciliary body and sclera. The sclera was glued to 8 separate PMMA segments and stretched radial by 5mm on an accommodation simulating lens stretching device. The point spread function was measured for each lens in the relaxed and stretched state for 5 human (ages 38-86 years) and 5 cynomolgus monkey (ages 53 - 67 months) fresh post mortem crystalline lenses. Results: Stretching induced measurable changes in the MTF. The cutoff frequency increased from 54.4±13.6 lp/mm unstretched to 59.5±21.4 lp/mm stretched in the post-presbyopic human and from 51.9±24.7 lp/mm unstretched to 57.7±18.5 lp/mm stretched cynomolgus monkey lenses. Conclusion: The results demonstrate the feasibility of measuring the optical quality of ex-vivo human and cynomolgus monkey lenses during simulated accommodation. Additional experiments are underway to quantify changes in optical quality induced by stretching.

Refractive Power and Biometric Properties of the Nonhuman Primate Isolated Crystalline Lens

Purpose. To characterize the age dependence of shape, refractive power, and refractive index of isolated lenses from nonhuman primates. Methods. Measurements were performed on ex vivo lenses from cynomolgus monkeys (cyno: n = 120; age, 2.7-14.3 years), rhesus monkeys (n = 61; age, 0.7-13.3 years), and hamadryas baboons (baboon: n = 16; age, 1.7-27.3 years). Lens thickness, diameter, and surface curvatures were measured with an optical comparator. Lens refractive power was measured with a custom optical system based on the Scheiner principle. The refractive contributions of the gradient, the surfaces, and the equivalent refractive index were calculated with optical ray-tracing software. The age dependence of the optical and biometric parameters was assessed. Results. Over the measured age range isolated lens thickness decreased (baboon: -0.04, cyno: -0.05, and rhesus: -0.06 mm/y) and equatorial diameter increased (logarithmically for the baboon and rhesus, and linearly for cyno: 0.07 mm/y). The isolated lens surfaces flattened and the corresponding refractive power from the surfaces decreased with age (-0.33, -0.48, and -0.68 D/y). The isolated lens equivalent refractive index decreased (only significant for the baboon, -0.001 D/y), and as a result the total isolated lens refractive power decreased with age (baboon: -1.26, cyno: -0.97, and rhesus: -1.76 D/y). Conclusions. The age-dependent trends in the optical and biometric properties, growth, and aging, of nonhuman primate lenses are similar to those of the pre-presbyopic human lens. As the lens ages, the decrease in refractive contributions from the gradient refractive index causes a rapid age-dependent decrease in maximally accommodated lens refractive power.

Biocompatibility of a nonpenetrating synthetic cornea in vascularized rabbit corneas

Purpose: This study was designed to assess feasibility and biocompatibility of a lamellar, nonperforating supraDescemetic Synthetic Cornea (sDSC) implanted in rabbit eyes after a corneal injury. Methods: Corneal vascularization and scarring was induced in the right eye of 15 rabbits by application of 1-heptanol and complete surgical removal of the limbus. An sDSC (7-mm diameter, 450-μm-thick optical zone, 100-μm-thick outer flange) was implanted after 45 ± 5 days. The keratoprostheses were implanted with their central optic part positioned on a completely exposed Descemets membrane (DM) while the outer flange was located in deep stroma. Three different materials were tested: hydrophobic PMMA (n = 5) and hydrophilic HEMA-MMA (n = 5) and HEMA-NVP (n = 5) with a water content of 34% and 75%, respectively. The corneal surface was covered with a conjunctiva-Tenon flap. Central flap trephination was performed after 63 ± 7 days. DM vascularization and scarring was assessed and graded after flap opening and weekly thereafter. Results: In all 15 consecutive cases implantation could be completed successfully without perforation of DM. Repair of the conjunctival flap had to be performed in five rabbits. Four months postoperatively, the flaps were opened. Four of five corneas (80%) with a PMMA implant and three of five (60%) with a HEMA-NVP75 implant had retained their original transparency. The others had developed significant neovascularization in the Descemet-sDSC optic interface. All corneas (100%) that received an sDSC made of HEMA-MMA34 displayed a completely clear DM without any vessels or scarring. DM was found firmly attached to the posterior surface of the optic. Conclusion: Implantation of a nonperforating synthetic cornea on top of an exposed DM is feasible. HEMA-MMA34 showed the most promising results. Because opening of the anterior chamber is not required, a lamellar supraDescemetic Synthetic Cornea would theoretically reduce some of the risks attributed to penetrating keratoprostheses.

Corneal stroma regeneration in felines after supradescemetic keratoprosthesis implantation.

Purpose: To show corneal regeneration in 3 cats that underwent lamellar keratectomy (90%) depth during supradescemetic keratoprosthetic implantation. Methods: Three 2-year-old cats that underwent spontaneous keratoprosthesis extrusion between 15 and 150 days after implanting a supradescemetic prosthesis into their right eyes were studied. Corneal structures and stroma thickness were evaluated by slit-lamp photographs, pachymetry, and confocal microscopy. Regenerated corneal epithelial cells, stroma matrix, and keratocyte morphology were studied with histology and transmission electron microscopy. Epithelial and stromal cell immunocharacterization was performed. Results: Corneas progressively regained normal thickness and improved clarity within 40 to 60 days. Slit-lamp photographs and pachymetry showed gains in stromal thickness until 600 μm or more. In vivo confocal microscopy showed the restoration of normal epithelium and stroma in all cats. Corneal nerves were seen in the regenerated stroma of 2 cats. Immunostaining showed absent α-smooth muscle actin (SMA) expression and a keratin K3-expressing epithelium. Electron microscopy showed regeneration of normal epithelium with a well-formed basement membrane, organized corneal lamellae, and the presence of normal keratocytes. Conclusion: Felines are capable of regenerating corneal structures including epithelium and reinnervated stroma matrix after deep lamellar keratectomy. The use of feline models in corneal keratoprosthesis is therefore questionable.

Validation of the barraquer tonometer for high intraocular pressure estimation

PURPOSE To compare the pneumatonometer and the Tono-Pen XL in a closed ex-vivo system in human eye bank eyes at high intraocular pressures (IOP) and evaluate the validity of high IOP measurements with the Barraquer tonometer. METHODS Intraocular pressure was monitored by cannulation of the anterior chamber and vitreous cavity in eight human cadaver eyes (mean donor age: 77.3 +/- 4.9 years, range: 72 to 84 years). Intraocular pressure measurements were taken at 50, 65, and 90 mmHg with the Tono-Pen XL and pneumatonometer. Intraocular pressure was raised to 110 mmHg and then the eyes were deflated slowly until they reached 50 mmHg. Pressure readings with the Barraquer tonometer were recorded when the corneal tonometer interface reached the inner and outer rings. RESULTS The Tono-Pen XL underestimated IOP, a tendency that was more evident at higher IOP In contrast, the pneumatonometer was more accurate and reliable at IOP of 50 and 65 mmHg but its readings underestimated IOP at 90 mmHg. The Barraquer tonometer used in this experiment accurately estimated high IOP A variability of 5.9 mmHg and 5.8 mmHg were recorded for the inner and outer ring, respectively. CONCLUSIONS The Tono-pen XL is an inadequate instrument to assess pressures normally encountered during LASIK flap creation in an ex vivo model using human cadaver eyes. The pneumatonometer and the Barraquer tonometer are accurate instruments at high IOP; however, the pneumatonometer underestimated pressures around 90 mmHg.

Treatment of retinal detachment with an encircling band and buckle implant: A comparative pilot study between poly (styrene- b -isobutylene- b -styrene) (SIBS) and trimethyl terminated polydimethylsiloxane (PDMS)

Purpose: To demonstrate the biocompatibility of SIBS implants as compared to PDMS implants in the treatment of retinal detachment in New Zealand White (NZW) rabbit model.1,2 Introduction: Scleral encircling bands, fixation rings and buckles are utilized for closure of retinal breaks and retina reattachment. The FDA approved PDMS-implant is associated with several post-operative complications, involving thick-fibrotic encapsulations. SIBS, an elastomeric triblock copolymer, was recently FDA approved for use in a cardiovascular drug eluting stent (TAXUSTM, Boston Scientific Corp., MA) and showed excellent biocompatibility and slow drug release capability. Materials and Methods: SIBS (9-mol%-styrene) implants were fabricated (InnFocus LLC, USA) to match PDMS implants (Labtician, Inc, Canada) dimensions. 5 NZW rabbits received SIBS and 4, PDMS-implants. Post-operative exam sequence: day 1 and 2, week 1, 2, 3, 4 and 6, and monthly thereafter for up to 1 year. Anatomohistopathology exams sequence: one SIBS animal at 6 weeks and one animal of each treatment group at 3 and 6-months, and two at 12-months. Results: SIBS compared to PDMS animals exhibited less inflammation and a better buckling effect during the first 6 weeks. At POD 9 months, the conjunctival injection in the SIBS rabbit was none as opposed to the PDMS value and the buckling effect for both groups were equal. There were no visible signs of encapsulation with SIBS. There were no infections in the 9 animals and none of the implants extruded thus far (<10 months). Conclusion: SIBS encircling bands, sleeves, and buckle implants are well tolerated in the rabbit model.

A novel orbital tissue expander (OTE): design, in vitro, and in vivo studies

Purpose: To assess the efficacy of a novel orbital tissue expander (OTE) in treating congenital anophthalmic and microphthalmic infants. Methods: The OTE implant is an inflatable (0.5 to >6cc) silicone rubber globe sliding on a titanium T-shaped bone plate secured to the temporal bone with 1mm titanium screws. In vitro testing was performed to assess injection volume versus diameter measurements to determine consistency between devices, flex fatigue for durability of the implants when compressed, weight change in isotonic saline at 37°C to mimic human body temperature, seal durability by puncturing the globe numerous times while inflating, capacity before rupture to determine the maximum amount of saline it is able to contain, and effective sterilization. Ex-vivo testing was performed for adjustments prior to in vivo study. An OTE was then implanted in five 2-week old kittens (OS only) and inflated in 0.5cc increments. Three control animals received enucleation alone. All 8 animals were followed for 18 weeks and underwent euthanasia for morphological and histopathological analysis. Results: In vitro testing confirmed a <5% diameter variance between different OTEs inflated in 0.5cc increments up to 5.0cc, <5% weight change in isotonic saline at 37°C over 7 weeks, <3% weight change over 14 months in the fatigue tester, and no quantifiable leakage (<1mg) after 65 consecutive 30ga needle punctures. The OTEs were successfully sterilized by autoclave and easily secured in the orbit of postmortem kittens. The in vivo study demonstrated biocompatibility of the implant which stimulates orbital bone growth resulting in almost no difference between the implanted socket and the control eye of the cat. There were no adverse effects in the normal maturation, weight gain, and food intake of the cats. Light microscopy showed no signs of foreign body reaction. Pictures of the implants were obtained by using a shadow-photogrammetry system to compare the explanted OTE with the OD control eye. Conclusion: In vitro and in vivo studies show the implants potential to safely treat anophthalmic and microphthalmic infants.

Non-contact optical measurement of lens capsule thickness during simulated accommodation

Purpose: To non-invasively measure the thickness of the anterior and posterior lens capsule, and to determine if it significantly changes during accommodation. Methods: Anterior and posterior capsule thickness was measured on post-mortem lenses using a non-contact optical system using a focus-detection technique. The optical system uses a 670nm laser beam delivered to a single-mode fiber coupler. The output of the fiber coupler is focused on the tissue surface using an aspheric lens (NA=0.68) mounted on a translation stage with a motorized actuator. Light reflected from the sample surface is collected by the fiber coupler and sent to a photoreceiver connected to a computer-controlled data acquisition system. Optical intensity peaks are detected when the aspheric lens is focused on the capsule boundaries. The capsule thickness is equal to the distance traveled between two peaks multiplied by the capsule refractive index. Anterior and posterior lens capsule thickness measurements were performed on 18 cynomolgus (age average: 6±1 years, range: 4-7 years) eyes, 1 rhesus (age: 2 years) eye, and 12 human (age average: 65±16, range: 47-92) eyes during simulated accommodation. The mounted sample was placed under the focusing objective of the optical system so that the light was incident on the center pole. Measurements were taken of the anterior lens capsule in the unstretched and the stretched 5mm states. The lens was flipped, and the same procedure was performed for the posterior lens capsule. Results: The precision of the optical system was determined to be ±0.5um. The resolution is 4um and the sensitivity is 52dB. The human anterior lens capsule thickness was 6.0±1.2um unstretched and 4.9±0.9um stretched (p=0.008). The human posterior lens capsule was 5.7±1.2um unstretched and 5.7±1.4um stretched (p=0.974). The monkey anterior lens capsule thickness was 5.9±1.9um unstretched and 4.8±1.0um stretched (p=0.002). The monkey posterior lens capsule was 5.9±2.0um unstretched and 5.1±1.3um stretched (p=0.128). Conclusions: The results indicate that the primate anterior lens capsule thickness changes during accommodation.

Endocapsular hyperthermia probe to prevent posterior capsular opacification

Purpose: To evaluate the feasibility to induce lens epithelial cell death with intraoperative hyperthermia for prevention of secondary cataract. Methods: A prototype miniature resistive hyperthermia probe was designed. The probe contained a thermocouple for temperature feed-back. A timer allowed monitoring of the electrical driving of the hyperthermia probe and the temperature induced as a function of time. To model the heating response, a simple model of the lens capsule was constructed using a thin acrylic plastic shell embedded in a sponge immersed in a water bath at 37°C. The shell was filled with sodium hyaluronate. The probe was positioned at the center of the shell with the thermocouple next to the wall. An experimental protocol was developed to assess the feasibility of increasing the temperature of the human lens to hyperthermia levels in fresh cadaver eyes: An annular metal ring was bonded just below the limbus, the cornea and iris were sectioned, the lens material was removed through a central 5mm diameter capsulorhexis, the capsule was filled with SHA and the globe was set on a temperature-controlled cylindrical vial. Preliminary Results: At 3.3W (2.2V, 1.5A) the shells content increases from 37°C to 51°C in 30s. At that temperature, LEC death is expected to occur within 1sec. Conclusion: This preliminary study demonstrates the feasibility of increasing the temperature of the capsular bag to kill LECs by hyperthermia.